WB Stripping SolutionFor Subsequent Detections after Western Blotting

Application

WB Stripping Solution removes conjugated antibodies from blots, enabling subsequent detections with different antigens on the very same blotting membrane. After the first antigen-antibody reaction and following chemiluminescent visualization, the antibody can be removed by the WB Stripping Solution. A second antigen-antibody reaction can be conducted on the same blotting membrane. The same blotting membrane can be probed 2-5 times if chemiluminescent detection is employed. Alternately, one blot can be used multiple times under different conditions to optimise results of antigen-antibody reactions!

Features

Caution

If antibody remains on the membrane, it can be detected after the new antigen-antibody reaction causing false result.

-Check whether any labeled antibody complex remains after the stripping by repeated chemiluminescence detection.

-Check whether any non-labeled (primary) antibody remains by chemiluminescence detection upon repeated reaction with the secondary antibody.

-Depending on the strength of the antigen-antibody reaction, the antibody complex may remain on the membrane after 5-15 minutes stripping treatment at room temperature. It is recom-mended to optimize the conditions beforehand. If antibodys remain, it is recommended to extend the stripping time (30 minutes to several hours) and/or raise temperature (37 to 60 oC). Perform blocking again if necessary.

-The product is an acidic solution (pH 2-3). Wear protective chemical-resistant clothes, gloves, and safety eye-wears. If the product accidentally comes into contact with skin, thoroughly wash the skin with water. Contact a physician if necessary.

-Use the product after it has been equilibrated at room tempera-ture.

-This product is not suitable for use for blotting membrane stained with TMB, DAB, 4-chloronaphtol or other visually detectable staining, because these stains will not disappear from the membrane.

-After use, the product shall be neutralized and discarded.-If the product is mixed with WB Stripping Solution Strong (Code No.:05677-65), a white precipitation will occur. When both products are used in turns, the blotting membrane must be washed throughly with a proper buffer or deionized water for 3-5 times. Use separate trays for different reagents.

Ordering informationCode No.05364-55

05677-65

Product nameWB Stripping Solution

WB Stripping Solution Strong

GradeSP

SP

StorageRT

R

PKG Size500 ml

500 ml

For research use only, not intended for diagnistic or drug use.

No heating: No odor:Fast:Ready to use:

reaction at room temperaturedoes not contain 2-mercaptoethanolstripping time 5-15 minutesone solution in one bottle

Data by Department of Bioscience and Biotechnology,Tokyo Institute of Technology.

1. First antigen-antibody reaction

BlockingWashPrimary abSecondary abDetection

2. Stripping↓

↓

Condition : RT, 15 min for conventional protocol

：Blocking One (30 min) (Code No.:03953-95) ：t-Tris Buffered Saline：anti-paxillin (mouse IgG)：anti-mouse IgG-POD：Chemiluminescence Detection Kit (commercially available product)

：Blocking One (30 min) (Code No.:03953-95) ：t-Tris Buffered Saline：anti-vinculine (mouse IgG) / anti-actin (mouse IgG)：anti-mouse IgG-POD：Chemi-Lumi One (Code No.:05027-20)

3. Second, differrent antigen-antibody reaction

BlockingWashPrimary abSecondary abDetection

3. Dispose stripping solution and wash the membrane with t-PBS or t-TBS for 5 minutes.4. Washed blotting membrane can be used for a second antigen -antibody reaction.

○

(Protocol)

Proof of antibody removal

Expiration

For blotting membrane (mini gel size), use 20 ml of the product.1. Wash chemiliminescent stain treated blotting membrane with t-PBS or t-TBS with 0.05% (v/v) Tween R 20 for 5 minutes.2. Pour enough WB stripping solution into the tray to cover the blotting membrane fully. Shake gently for 5-15 minutes.

Protocol

One year from manufacturing. Expiration date is stated on the product label (Exp. yy / mm)

For blotting membrane (mini gel size), use 20 ml of the product.All equipments (tray, tweezers etc) coming into contact with this product should be made of plastic.1. Wash chemiliminescent stain treated blotting membrane with deionized water for 1 minutes.2. Pour enough WB stripping solution into the tray to cover the blotting membrane fully. Shake gently for 5-15 minutes.3. Dispose stripping solution and wash the membrane with t-PBS or t-TBS with 0.05%(v/v) Tween R 20 for 5 minutes.4. Washed blotting membrane can be used for a second antigen -antibody reaction.

Western Blotting Solution StrongProtocol Expiration

One year from manufacturing. Expiration date is stated on the product label (Exp. yy / mm)

Ordering Information

For research use only, not intended for diagnistic or drug use.

NACALAI TESQUE, INC.Nijo Karasuma, Nakagyo-ku, Kyoto 604-0855, JAPANTEL: +81-75-251-1730 FAX: +81-75-251-1763Web site: http://www.nacalai.comE-mail: info.intl@nacalai.co.jp

○

If antibody remains on the membrane, it can be detected after the new antigen-antibody reaction causing false result.

-Check whether any labeled antibody complex remains after the stripping by repeated chemiluminescence detection.

-Check whether any non-labeled (primary) antibody remains by chemiluminescence detection upon repeated reaction with the secondary antibody.

Proof of antibody removal

Caution-All equipments (tray, tweezers etc) coming into contact with this product should be made of plastic. Contact with metal equipment compromise the performance of the product.

-Depending on the strength of the antigen-antibody reaction, the antibody complex may remain on the membrane after 5-15 minutes stripping treatment at room temperature. It is recom-mended to optimize the conditions beforehand. If antigens re-main, it is recommended to extend the stripping time (30 mi-nutes to several hours) and/or raise temperature (37 to 60 oC). Perform blocking again if necessary.

-The product has reductive property. Wear protecting chemical-resistant clothes, gloves, and safety eye-wears. If the product accidentally comes into contact with skin, thoroughly wash the skin with water. Contact a physician if necessary.

-Use the product after it has been equilibrated at room temper-ature.

-The product sould be stored at 2-8 oC. During long storage, the surfactant may precipitate. Precipitation causes change in the concentration. Therefore use the product after the precipitation has been dissolved.

-This product is not suitable for use for blotting membrane stained with TMB, DAB, 4-chloronaphtol or other visually de-tectable staining, because these stains will not disappear from the membrane.

-If the product is mixed with WB Stripping Solution (Code No.:05364-55), a white precipitation will occur. When both prod-ucts are used in turns, the blotting membrane must be washed throughly with a proper buffer or deionized water for 3-5 times. Use separate trays for different reagents.

Code No.

05677-65

Product name

WB Stripping Solution Strong

Grade

SP

Storage

R

PKG Size

500 ml